|Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism|
|Jiao-Yu Deng; Xian-En Zhang; Yuan Mang; Zhi-Ping Zhang; Ya-Feng Zhou; Qiang Liu; Hong-Bing Lu, Zhi-Jun Fu; Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China
|Source Publication||BIOSENSORS & BIOELECTRONICS
|Abstract||An oligonucleotide ligation assay-based DNA chip has been developed to detect single nucleotide polymorphism. Synthesized nonamers, complementary to the flanking sequences of the mutation sites in target DNA, were immobilized onto glass slides through disulfide bonds on their 5' terminus. Allele-specific pentamers annealed adjacent to the nonamers on the complementary target DNA, containing 5'-phosphate groups and biotin labeled 3'-ends, were mixed with the target DNA in tube. Ligation reactions between nonamers and pentamers were carried out on chips in the presence of T4 DNA ligase. Ligation products were directly visualized on chips through enzyme-linked assay. The effect of G:T mismatch at different positions of pentamers on the ligation were evaluated. The results showed that any mismatch between pentamer and the target DNA could lead to the decrease of ligation, which can be detected easily. The established approach was further used for multiplex detection of mutations in rpoB gene of rifampin-resistant Mycobacterium tuberculosis clinical isolates. (C) 2003 Elsevier B.V. All rights reserved.|
|Keyword||Oligonucleotide Ligation Assay
Single Nucleotide Polymorphism Detection
T4 Dna Ligase
|Department||Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China; Chinese Acad Sci, Wuhan Inst Hydrobiol, Wuhan 430072, Peoples R China; Wuhan TB Prevent & Cure Inst, Wuhan 430030, Peoples R China
; Biotechnology & Applied Microbiology
; Nanoscience & Nanotechnology
|Corresponding Author||Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China|
Jiao-Yu Deng,Xian-En Zhang,Yuan Mang,et al. Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism[J]. BIOSENSORS & BIOELECTRONICS,2004,19(10):1277-1283.
Jiao-Yu Deng.,Xian-En Zhang.,Yuan Mang.,Zhi-Ping Zhang.,Ya-Feng Zhou.,...&Zhang, XE, Chinese Acad Sci, Wuhan Inst Virol, Wuhan 430071, Peoples R China.(2004).Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism.BIOSENSORS & BIOELECTRONICS,19(10),1277-1283.
Jiao-Yu Deng,et al."Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism".BIOSENSORS & BIOELECTRONICS 19.10(2004):1277-1283.
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